BACKGROUND:In vitro screening and amplification are important links to harvest muscle-derived stem cells that are satisfactory to clinical requirement.OBJECTIVE:To probe into the method of isolation,culture and purification of skeletal muscle-derived stem cells from adult rats in vitro.METHODS:The skeletal muscle was obtained sterilely following adult Sprague Dawley rats were anesthetized.Muscle-derived stem cells were harvested using enzyme digestion with Ⅺ collagenase,Dispese and trypsogen,and then purified by Percoll density gradient centrifugation and differential adhesion method.Growth curves were recorded and MTT colorimetric technique was used to describe the effects of various kinds of inoculum density on cell growth.Cells were identified by immunocytochemistry.RESULTS AND CONCLUSION:Primary muscle-derived stem cells were less in volume,lower adherence and well refraction,appearing as globular or fusiform or spindle and slowly multiplication.Following subculture,complete medium containing 20%serum was added.Cell number was greatest when cell density was 1×109/L,which was the optimal density.Cells at passages 1-4 grew well.Cells showed desmin(+),CD34(+),CD45(-)and Sca-1(+)by immunocytochemistry.Results verified that high-purity muscle-derived stem cells can be obtained in vitro and amplified successfully following primary culture.

Objectives: To evaluate the clinical significance of nuclear matrix protein 22 (NMP 22) in the detection of bladder transitional cell carcinoma (BTCC) and compare with voided urine cytology(VUC). Methods: A total of 69 cases with voided urine samples for NMP 22 and VUC test were included in this study. Thirty of them were BTCC patients(BTCC group) and twenty nine suffered from other urological diseases (nonbladder cancer group, NBC group). Ten were healthy volunteers (control group). Results: The NMP 22 values for BTCC group (67.3 U/ml) were significantly higher than that of NBC group(7.4 U/ml) and control group (4.3 U/ml)( P 0.05). NMP 22 was more sensitive than VUC in low grade BTCC(Ⅰ,Ⅱ)(62.50% vs 12.50%,P 0.05). Conclusions:Urinary NMP 22 is a useful marker for the early diagnosis of BTCC. It is more sensitive than VUC in low stage and grade BTCC.

Objective To evaluate the effectiveness and safety of flexible ureteroscopy lithotripsy (FURL) combined with holmium laser for the concurrent treatment of bilateral upper ureteral tract calculi.Methods The clinical data in 43 patients with bilateral upper ureteral tract calculi concurrently treated by FURL combined with holmium laser in our hospital from September 2014 to November 2016 were retrospectively analyzed.All cases conducted CT scanning before operation.The flexible ureteroscope was intraoperatively placed and the lithotripsy was conducted by using 200 μm optical fiber at a power of 0.8-1.0 J/10-20Hz,which was coordinated by reticular basket.After lithotripsy,bilateral urethral stents were placed.KUB or CT was re-examined on postoperative 1 d.The cases of residual stone were performed KUB again after removing urethral stent after 4-6 weeks.Results All cases were successfully placed the flexible ureteroscope by one time.The mean operation time was (101.5±37.2) min and the overall stone-free rate (SFR) was 81.4 ％ (35/43).SFR was 100％ in the patients with the stone burden less than 30 mm and 63.2％ in the patients with the stone burden ≥30 mm,the difference was statistically significant(P＜0.05).One case appeared fever (39.5 ℃) after operation,1 case developed subcapsular renal hematoma and no other severe complications occurred.Conclusion FURL combined with holmium laser is an effective means to concurrently treating bilateral upper ureteral calculi,especially for the patients with stone burden ＜30 mm,which has higher stone clearance rate with good safety.